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Cloning, nucleotide sequence, and expression of the gene encoding a novel dioxygenase involved in metabolism of carboxydiphenyl ethers in Pseudomonas pseudoalcaligenes POB310

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Abstract

Pseudomonas pseudoalcaligenes strain POB310 degrades 3-and 4-carboxydiphenyl ether. The initial reaction involves an angular dioxygenation yielding an unstable hemiacetal that spontaneously decays to phenol and protocatechuate. We cloned a DNA fragment containing the gene encoding the initial, dioxygenase from an unstable, self-transmissible plasmid. Sequence analysis revealed two open reading frames encoding proteins with putative molecular masses of 46.3 and 33.6 kDa. The deduced amino acid sequences showed homologies to oxygenase and reductase subunits of aromatic ring-activating dioxygenases, and contained regions identical to consensus sequences that bind chloroplast-like and Rieske-type [2Fe2S] clusters suggesting that the initial dioxygenase is a class IA aromatic ring-activating dioxygenase system. Intitial dioxygenase activity was induced in bacteria grown in M9 minimal medium containing 3-or 40-carboxydiphenyl ether or phenol as carbon source, indicating that the regulation is dependent on the phenol pathway. The maximal specific activity was measured at the beginning of the exponential growth phase.

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Author notes

  1. Uwe Dehmel

    Present address: Department of Human and Animal Cell Cultures, DSM, German Collection of Microorganisms and Cell Cultures, Mascheroder Weg 1b, D-38124, Braunschweig, Germany

  2. Daryl F. Dwyer

    Present address: Department of Civil Engineering, University of Minnesota, 500 Pillsbury Drive, S. E., 55455, Minneapolis, MN, USA

  3. Karl-Heinrich Engesser

    Present address: ISWA, Abteilung Biologische Abluftreinigung, University of Stuttgart, Im Bandtäle 2, D-70569, Stuttgart, Germany

Authors and Affiliations

  1. Department of Microbiology, G.B.F. National Research Center for Biotechnology, Mascheroder Weg 1, D-38124, Braunschweig, Germany

    Uwe Dehmel, Kenneth N. Timmis & Daryl F. Dwyer

  2. Department of Microbiology, University of Stuttgart, Allmand Ring 31, D-70569, Stuttgart, Germany

    Karl-Heinrich Engesser

Authors
  1. Uwe Dehmel
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  2. Karl-Heinrich Engesser
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  3. Kenneth N. Timmis
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  4. Daryl F. Dwyer
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Correspondence to Daryl F. Dwyer.

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Dehmel, U., Engesser, KH., Timmis, K.N. et al. Cloning, nucleotide sequence, and expression of the gene encoding a novel dioxygenase involved in metabolism of carboxydiphenyl ethers in Pseudomonas pseudoalcaligenes POB310. Arch. Microbiol. 163, 35–41 (1995). https://doi.org/10.1007/BF00262201

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  • DOI: https://doi.org/10.1007/BF00262201

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